Brief exposure to small molecules allows induction of mouse embryonic fibroblasts into neural crest-like precursors

Yuzo Takayama; Tamami Wakabayashi; Hiroko Kushige; Yutaka Saito; Yoichiro Shibuya; Shinsuke Shibata; Wado Akamatsu; Hideyuki Okano; Yasuyuki S Kida

doi:10.1002/1873-3468.12572

Brief exposure to small molecules allows induction of mouse embryonic fibroblasts into neural crest-like precursors

FEBS Lett. 2017 Feb;591(4):590-602. doi: 10.1002/1873-3468.12572. Epub 2017 Feb 9.

Authors

Yuzo Takayama¹, Tamami Wakabayashi¹, Hiroko Kushige¹, Yutaka Saito^{2

3}, Yoichiro Shibuya^{1

4}, Shinsuke Shibata⁵, Wado Akamatsu⁶, Hideyuki Okano⁵, Yasuyuki S Kida¹

Affiliations

¹ Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan.
² Artificial Intelligence Research Center, National Institute of Advanced Industrial Science and Technology (AIST), Koto-ku, Tokyo, Japan.
³ Computational Bio Big-Data Open Innovation Laboratory (CBBD-OIL), National Institute of Advanced Industrial Science and Technology (AIST), Shinjuku-ku, Tokyo, Japan.
⁴ Department of Plastic and Reconstructive Surgery, University of Tsukuba, Ibaraki, Japan.
⁵ Department of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
⁶ Center for Genomic and Regenerative Medicine, Juntendo University, Bunkyo-ku, Tokyo, Japan.

Abstract

In this study, we propose a novel method for inducing neuronal cells by briefly exposing them to small-molecule cocktails in a step-by-step manner. Global gene expression analysis with immunohistochemical staining and calcium flux assays reveal the generation of neurons from mouse embryonic fibroblasts. In addition, time-lapse imaging of neural precursor-specific enhancer expression and global gene expression analyses show that the neurons are generated by passing through a neural crest-like precursor stage. Consistent with these results, the neural crest-like cells are able to differentiate into neural crest lineage cells, such as sympathetic neurons, adipocytes, osteocytes, and smooth muscle cells. Therefore, these results indicate that brief exposure to chemical compounds could expand and induce a substantial multipotent cell population without viral transduction.

Keywords: neural crest; peripheral neuron; small molecules.

MeSH terms

Adipocytes / drug effects
Adipocytes / metabolism
Animals
Cell Differentiation / drug effects*
Cell Differentiation / genetics
Cells, Cultured
Colforsin / pharmacology
Embryo, Mammalian / cytology
Fibroblasts / cytology
Fibroblasts / drug effects*
Fibroblasts / metabolism
Gene Expression Profiling / methods
Gene Ontology
Immunohistochemistry
Mice, Inbred C57BL
Microtubule-Associated Proteins / genetics
Microtubule-Associated Proteins / metabolism
Myocytes, Smooth Muscle / drug effects
Myocytes, Smooth Muscle / metabolism
Neural Crest / cytology
Neural Crest / drug effects*
Neural Crest / metabolism
Neural Stem Cells / cytology
Neural Stem Cells / drug effects*
Neural Stem Cells / metabolism
Neurons / drug effects
Neurons / metabolism
Osteocytes / drug effects
Osteocytes / metabolism
Pyridines / pharmacology
Pyrimidines / pharmacology
Reverse Transcriptase Polymerase Chain Reaction
Small Molecule Libraries / pharmacology*
Time-Lapse Imaging / methods
Tranylcypromine / pharmacology
Valproic Acid / pharmacology

Substances

Chir 99021
Microtubule-Associated Proteins
Mtap2 protein, mouse
Pyridines
Pyrimidines
Small Molecule Libraries
Colforsin
Tranylcypromine
Valproic Acid

Associated data

GENBANK/GSE78890